Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

Casper Wilkens; Kyle D. Auger; Nolan T. Anderson; David A. Meekins; Madushi Raththagala; Maher  Abou Hachem; Christina M. Payne; Matthew S. Gentry; Birte Svensson

doi:10.1002/1873-3468.12027

Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

Casper Wilkens, Kyle D. Auger, Nolan T. Anderson, David A. Meekins, Madushi Raththagala, Maher Abou Hachem, Christina M. Payne, Matthew S. Gentry, Birte Svensson

University of Kentucky

Research output: Contribution to journal › Journal article › Research › peer-review

Abstract

The plant glucan phosphatases Starch EXcess 4 (SEX4) and Like Sex Four2 (LSF2) apply different starch binding mechanisms. SEX4 contains a carbohydrate binding module, and LSF2 has two surface binding sites (SBSs). We determined K_Dapp for amylopectin and amylose, and KD for β-cyclodextrin and validated binding site mutants deploying affinity gel electrophoresis (AGE) and surface plasmon resonance. SEX4 has a higher affinity for amylopectin; LSF2 prefers amylose and β-cyclodextrin. SEX4 has 50-fold lower K_Dapp for amylopectin compared to LSF2. Molecular dynamics simulations and AGE data both support long-distance mutual effects of binding at SBSs and the active site in LSF2.

Original language	English
Journal	FEBS Letters
Volume	590
Issue number	1
Pages (from-to)	118-128
Number of pages	11
ISSN	0014-5793
DOIs	https://doi.org/10.1002/1873-3468.12027
Publication status	Published - 2016

Keywords

Affinity gel electrophoresis
Carbohydrate binding domain
Like Sex Four2
Starch Excess 4
Surface binding sites
Surface plasmon resonance

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10.1002/1873-3468.12027

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@article{c9941911c21d44988509a8444fd56a22,

title = "Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose",

abstract = "The plant glucan phosphatases Starch EXcess 4 (SEX4) and Like Sex Four2 (LSF2) apply different starch binding mechanisms. SEX4 contains a carbohydrate binding module, and LSF2 has two surface binding sites (SBSs). We determined KDapp for amylopectin and amylose, and KD for β-cyclodextrin and validated binding site mutants deploying affinity gel electrophoresis (AGE) and surface plasmon resonance. SEX4 has a higher affinity for amylopectin; LSF2 prefers amylose and β-cyclodextrin. SEX4 has 50-fold lower KDapp for amylopectin compared to LSF2. Molecular dynamics simulations and AGE data both support long-distance mutual effects of binding at SBSs and the active site in LSF2.",

keywords = "Affinity gel electrophoresis, Carbohydrate binding domain, Like Sex Four2, Starch Excess 4, Surface binding sites, Surface plasmon resonance",

author = "Casper Wilkens and Auger, {Kyle D.} and Anderson, {Nolan T.} and Meekins, {David A.} and Madushi Raththagala and {Abou Hachem}, Maher and Payne, {Christina M.} and Gentry, {Matthew S.} and Birte Svensson",

year = "2016",

doi = "10.1002/1873-3468.12027",

language = "English",

volume = "590",

pages = "118--128",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "JohnWiley & Sons Ltd.",

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TY - JOUR

T1 - Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

AU - Wilkens, Casper

AU - Auger, Kyle D.

AU - Anderson, Nolan T.

AU - Meekins, David A.

AU - Raththagala, Madushi

AU - Abou Hachem, Maher

AU - Payne, Christina M.

AU - Gentry, Matthew S.

AU - Svensson, Birte

PY - 2016

Y1 - 2016

N2 - The plant glucan phosphatases Starch EXcess 4 (SEX4) and Like Sex Four2 (LSF2) apply different starch binding mechanisms. SEX4 contains a carbohydrate binding module, and LSF2 has two surface binding sites (SBSs). We determined KDapp for amylopectin and amylose, and KD for β-cyclodextrin and validated binding site mutants deploying affinity gel electrophoresis (AGE) and surface plasmon resonance. SEX4 has a higher affinity for amylopectin; LSF2 prefers amylose and β-cyclodextrin. SEX4 has 50-fold lower KDapp for amylopectin compared to LSF2. Molecular dynamics simulations and AGE data both support long-distance mutual effects of binding at SBSs and the active site in LSF2.

AB - The plant glucan phosphatases Starch EXcess 4 (SEX4) and Like Sex Four2 (LSF2) apply different starch binding mechanisms. SEX4 contains a carbohydrate binding module, and LSF2 has two surface binding sites (SBSs). We determined KDapp for amylopectin and amylose, and KD for β-cyclodextrin and validated binding site mutants deploying affinity gel electrophoresis (AGE) and surface plasmon resonance. SEX4 has a higher affinity for amylopectin; LSF2 prefers amylose and β-cyclodextrin. SEX4 has 50-fold lower KDapp for amylopectin compared to LSF2. Molecular dynamics simulations and AGE data both support long-distance mutual effects of binding at SBSs and the active site in LSF2.

KW - Affinity gel electrophoresis

KW - Carbohydrate binding domain

KW - Like Sex Four2

KW - Starch Excess 4

KW - Surface binding sites

KW - Surface plasmon resonance

U2 - 10.1002/1873-3468.12027

DO - 10.1002/1873-3468.12027

M3 - Journal article

C2 - 26763114

SN - 0014-5793

VL - 590

SP - 118

EP - 128

JO - FEBS Letters

JF - FEBS Letters

IS - 1

ER -

Plant α-glucan phosphatases SEX4 and LSF2 display different affinity for amylopectin and amylose

Abstract

Keywords

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